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Updated: Apr 10, 2026
Mitochondrial Preparation from Microglia for Glycan Analysis
Published on: May 30, 2025
ADT-OH promotes mitophagy and suppresses the cytosolic mtDNA-cGAS-STING inflammatory cascade in microglia
Xiao-Ou Hou1,2, Miao Wang2, Rong Deng1,2
1Department of Neurology and Clinical Research Center of Neurological Disease, The Second Affiliated Hospital of Soochow University, Suzhou, 215004, China.
Abstract:
Mitochondrial dysfunction, driven by genetic susceptibility or environmental insults, contributes to the pathogenesis of neurodegenerative disorders, including Parkinson's disease (PD). Mitophagy is a selective pathway that eliminates dysfunctional mitochondria, and mitophagy inducers hold therapeutic promise for neurodegeneration. However, the arsenal of specific, clinically viable inducers remains limited. ADT-OH, a slow-release H2S compound, was recently reported to induce mitochondrial uncoupling through sulfide-quinone oxidoreductase (SQR)-mediated oxidation of H2S. In this study, we report that ADT-OH elicits mitophagic flux in microglia. This is evidenced by the reduced steady-state levels of mitochondrial marker proteins (TOM20, COXIV, and HSP60), enhanced mitochondrial fission dynamics, and mitochondrial translocation into lysosomes, as visualized by the mt-Keima probe. Mechanistically, its mitophagy-promoting effect is dependent on SQR-mediated mitochondrial uncoupling and subsequent activation of PINK1-PARKIN signaling. Importantly, ADT-OH abrogates the accumulation of dysfunctional mitochondria and the subsequent cytosolic release of mitochondrial DNA in α-synuclein preformed fibrils (α-Syn PFF)-challenged microglia, thereby blunting the activation of the cGAS-STING pathway and the downstream production of inflammatory mediators. Furthermore, systemic administration of ADT-OH dampened microglial activation and cGAS expression in α-Syn-overexpressing PD mice, thereby mitigating the loss of midbrain dopaminergic neurons and ameliorating motor coordination deficits. Collectively, our findings demonstrate that ADT-OH exerts robust neuroprotective effects in PD models, both in vitro and in vivo, by enhancing mitophagy and inhibiting microglia-mediated neuroinflammation.
Insights
ADT-OH, a novel mitophagy inducer, protects against Parkinson's disease by clearing dysfunctional mitochondria in microglia. This reduces neuroinflammation and preserves dopaminergic neurons, offering therapeutic potential for neurodegeneration.
Area of Science:
- Neuroscience
- Cell Biology
- Pharmacology
Background:
- Mitochondrial dysfunction is a key factor in neurodegenerative diseases like Parkinson's disease (PD).
- Mitophagy, the process of clearing damaged mitochondria, is a potential therapeutic target for neurodegeneration.
- Existing mitophagy inducers are limited, highlighting the need for new agents.
Purpose of the Study:
- To investigate the potential of ADT-OH, a hydrogen sulfide (H2S) releasing compound, as a mitophagy inducer in microglia.
- To elucidate the mechanism by which ADT-OH induces mitophagy and its downstream effects on neuroinflammation.
- To evaluate the therapeutic efficacy of ADT-OH in preclinical models of Parkinson's disease.
Main Methods:
- Assessed mitophagy flux in microglia using mitochondrial marker proteins and live-cell imaging with mt-Keima.
- Investigated the role of sulfide-quinone oxidoreductase (SQR) and PINK1-PARKIN signaling in ADT-OH-induced mitophagy.
- Examined the impact of ADT-OH on mitochondrial dysfunction, mitochondrial DNA release, and cGAS-STING pathway activation in microglia challenged with alpha-synuclein.
- Evaluated the in vivo efficacy of ADT-OH in an alpha-synuclein overexpression mouse model of Parkinson's disease.
Main Results:
- ADT-OH treatment increased mitophagic flux in microglia, evidenced by reduced mitochondrial proteins, increased fission, and lysosomal engulfment.
- The mitophagy-promoting effect of ADT-OH was dependent on SQR-mediated mitochondrial uncoupling and PINK1-PARKIN activation.
- ADT-OH prevented mitochondrial dysfunction and DNA release in alpha-synuclein-challenged microglia, suppressing cGAS-STING pathway activation and neuroinflammation.
- Systemic ADT-OH administration reduced microglial activation, protected dopaminergic neurons, and improved motor deficits in a PD mouse model.
Conclusions:
- ADT-OH effectively induces mitophagy in microglia via SQR-dependent mitochondrial uncoupling and PINK1-PARKIN signaling.
- ADT-OH mitigates alpha-synuclein-induced neuroinflammation by inhibiting mitochondrial dysfunction and the cGAS-STING pathway.
- ADT-OH demonstrates significant neuroprotective effects in vitro and in vivo, positioning it as a promising therapeutic candidate for Parkinson's disease.
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